Metabolomics of Mammalian and Cellular Models of Aging

Aging is often associated with impaired cognition and a progressive loss of organ function over time accompanied by an increased susceptibility for many disorders, including Alzheimer\u27s disease (AD), Parkinson\u27s disease (PD), heart disease, osteoporosis, type II diabetes, and many forms of cancer. With a rapidly aging population, the negative impacts of aging and age-related disorders is a major cause of increased human suffering both for affected individuals and for families and caregivers. Metabolic changes are also apparent in normal aging, but may increase in magnitude or nature with accompanying disease states or with accelerated aging. Thus, studying aging in a disease state, or in a disorder characterized by accelerated aging, will facilitate identification of these changes. Trisomy of chromosome 21 (HSA21), or Down syndrome (DS), is an intellectual disability characterized by premature aging. We hypothesize that trisomy causes disruption of the metabolome leading to an accelerated aging phenotype. In the Ts65Dn mouse model of DS, a premature aging phenotype is also observed along with other common comorbidities associated with human DS. Here, we report changes in the both global and targeted metabolomics (the study of small molecules) in the brains of the Ts65Dn mouse. We also report that long-term treatment with microencapsulated dietary rapamycin changes the metabolomic profiles in a manner consistent with increases in healthspan. Purines are molecules essential for many cell processes, including RNA and DNA synthesis, regulation of enzyme activity, protein synthesis and function, energy metabolism and transfer, essential coenzyme function, and cell signaling. Purines are produced via the de novo purine biosynthesis pathway. Mutations in purine biosynthetic genes can lead to developmental anomalies in lower vertebrates. Alterations in PAICS (phosphoribosyl- aminoimidazole carboxylase/phosphoribosylaminoimidazole succinocarboxamide synthetase) expression in humans have been associated with various types of cancer. Mutations in adenylosuccinate lyase (ADSL, E.C. 4.3.2.2) or 5-aminoimidazole-4-carboxamide ribonucleotide formyl-transferase/IMP cyclohydrolase (ATIC, E.C. 2.1.2.3/E.C. 3.5.4.10) lead to inborn errors of metabolism with a range of clinical symptoms, including developmental delay, severe neurological symptoms, renal stones, combined immunodeficiency, and autistic features. The pathogenetic mechanism is unknown for any of these conditions, and no effective treatments exist. The study of cells carrying mutations in the various de novo purine biosynthesis pathway genes provides one approach to analysis of purine disorders. Here we report the characterization of AdeD Chinese hamster ovary (CHO) cells, which carry genetic mutations encoding p.E177K and p.W363* variants of PAICS. Both mutations impact PAICS structure and completely abolish its biosynthesis. Additionally, we describe a sensitive and rapid analytical method for detection of purine de novo biosynthesis intermediates based on high performance liquid chromatography with electrochemical detection. Using this technique, we detected accumulation of 5-Aminoimidazole riboside (AIR) in AdeD cells. In AdeI cells, mutant for the ADSL gene, we detected accumulation of Succinylaminoimidazole carboxamide riboside (SAICAR) and adenylosuccinate (SAMP) and, somewhat unexpectedly, accumulation of AIR. The use of HPLC coupled electrochemical detection in combination with cellular assay methods have great potential for metabolite profiling of de novo purine biosynthesis pathway mutants, identification of novel genetic defects of purine metabolism in humans, and elucidating the regulation of this critical metabolic pathway

The Use of Mouse Models for Understanding the Biology of Down Syndrome and Aging

Down syndrome is a complex condition caused by trisomy of human chromosome 21. The biology of aging may be different in individuals with Down syndrome; this is not well understood in any organism. Because of its complexity, many aspects of Down syndrome must be studied either in humans or in animal models. Studies in humans are essential but are limited for ethical and practical reasons. Fortunately, genetically altered mice can serve as extremely useful models of Down syndrome, and progress in their production and analysis has been remarkable. Here, we describe various mouse models that have been used to study Down syndrome. We focus on segmental trisomies of mouse chromosome regions syntenic to human chromosome 21, mice in which individual genes have been introduced, or mice in which genes have been silenced by targeted mutagenesis. We selected a limited number of genes for which considerable evidence links them to aspects of Down syndrome, and about which much is known regarding their function. We focused on genes important for brain and cognitive function, and for the altered cancer spectrum seen in individuals with Down syndrome. We conclude with observations on the usefulness of mouse models and speculation on future directions

Rapamycin Treatment Ameliorates Age-Related Accumulation of Toxic Metabolic Intermediates in Brains of the Ts65Dn Mouse Model of Down Syndrome and Aging

Down syndrome (DS), caused by trisomy of chromosome 21, is the most common genetic cause of intellectual disability. Individuals with DS exhibit changes in neurochemistry and neuroanatomy that worsen with age, neurological delay in learning and memory, and predisposition to Alzheimer’s disease. The Ts65Dn mouse is the best characterized model of DS and has many features reminiscent of DS, including developmental anomalies and age-related neurodegeneration. The mouse carries a partial triplication of mouse chromosome 16 containing roughly 100 genes syntenic to human chromosome 21 genes. We hypothesized that there would be differences in brain metabolites with trisomy and age, and that long-term treatment with rapamycin, mechanistic target of rapamycin (mTOR) inhibitor and immunosuppressant, would correct these differences. Using HPLC coupled with electrochemical detection, we identified differences in levels of metabolites involved in dopaminergic, serotonergic, and kynurenine pathways in trisomic mice that are exacerbated with age. These include homovanillic acid, norepinephrine, and kynurenine. In addition, we demonstrate that prolonged treatment with rapamycin reduces accumulation of toxic metabolites (such as 6-hydroxymelatonin and 3-hydroxykynurenine) in aged mice

The Great Escape: The Role of Self-esteem and Self-related Cognition in Terror Management

Integrating terror management theory and objective self-awareness theory, we propose the existential escape hypothesis, which states that people with low self-esteem should be especially prone to escaping self-awareness as a distal response to thoughts of death. This is because they lack the means to bolster the self as a defense, and the propensity to bolster the self reduces the motivation to escape from self-awareness. Five studies supported this hypothesis. Individuals low, but not high, in self-esteem scored lower on a measure of private self-awareness (Study 1), showed less implicit self-activation (Studies 2 & 3), were more likely to choose to write about others than themselves (Study 4), and consumed more alcohol in a field study at a nightclub (Study 5) in response to mortality reminders. Implications for terror management theory (highlighting an additional route to defend against mortality awareness), self-regulation, physical health and well-being are discussed

The Global Evolution of Giant Molecular Clouds II: The Role of Accretion

We present virial models for the global evolution of giant molecular clouds. Focusing on the presence of an accretion flow, and accounting for the amount of mass, momentum, and energy supplied by accretion and star formation feedback, we are able to follow the growth, evolution, and dispersal of individual giant molecular clouds. Our model clouds reproduce the scaling relations observed in both galactic and extragalactic clouds. We find that accretion and star formation contribute contribute roughly equal amounts of turbulent kinetic energy over the lifetime of the cloud. Clouds attain virial equilibrium and grow in such a way as to maintain roughly constant surface densities, with typical surface densities of order 50 - 200 Msun pc^-2, in good agreement with observations of giant molecular clouds in the Milky Way and nearby external galaxies. We find that as clouds grow, their velocity dispersion and radius must also increase, implying that the linewidth-size relation constitutes an age sequence. Lastly, we compare our models to observations of giant molecular clouds and associated young star clusters in the LMC and find good agreement between our model clouds and the observed relationship between H ii regions, young star clusters, and giant molecular clouds.Comment: 23 Pages, 9 Figures. Accepted to Ap

Reduced purine biosynthesis in humans after their divergence from Neandertals

We analyze the metabolomes of humans, chimpanzees, and macaques in muscle, kidney and three different regions of the brain. Although several compounds in amino acid metabolism occur at either higher or lower concentrations in humans than in the other primates, metabolites downstream of adenylosuccinate lyase, which catalyzes two reactions in purine synthesis, occur at lower concentrations in humans. This enzyme carries an amino acid substitution that is present in all humans today but absent in Neandertals. By introducing the modern human substitution into the genomes of mice, as well as the ancestral, Neandertal-like substitution into the genomes of human cells, we show that this amino acid substitution contributes to much or all of the reduction of de novo synthesis of purines in humans

Molecular and atomic gas in dust lane early-type galaxies - I : Low star-formation efficiencies in minor merger remnants

In this work we present IRAM-30m telescope observations of a sample of bulge-dominated galaxies with large dust lanes, which have had a recent minor merger. We find these galaxies are very gas rich, with H2 masses between 4x10^8 and 2x10^10 Msun. We use these molecular gas masses, combined with atomic gas masses from an accompanying paper, to calculate gas-to-dust and gas-to-stellar mass ratios. The gas-to-dust ratios of our sample objects vary widely (between ~50 and 750), suggesting many objects have low gas-phase metallicities, and thus that the gas has been accreted through a recent merger with a lower mass companion. We calculate the implied minor companion masses and gas fractions, finding a median predicted stellar mass ratio of ~40:1. The minor companion likely had masses between ~10^7 - 10^10 Msun. The implied merger mass ratios are consistent with the expectation for low redshift gas-rich mergers from simulations. We then go on to present evidence that (no matter which star-formation rate indicator is used) our sample objects have very low star-formation efficiencies (star-formation rate per unit gas mass), lower even than the early-type galaxies from ATLAS3D which already show a suppression. This suggests that minor mergers can actually suppress star-formation activity. We discuss mechanisms that could cause such a suppression, include dynamical effects induced by the minor merger.Peer reviewe

Treatment of Community-Acquired Pneumonia in Immunocompromised Adults:A Consensus Statement Regarding Initial Strategies

Background Community-acquired pneumonia (CAP) guidelines have improved the treatment and outcomes of patients with CAP, primarily by standardization of initial empirical therapy. But current society-published guidelines exclude immunocompromised patients. Research Question There is no consensus regarding the initial treatment of immunocompromised patients with suspected CAP. Study Design and Methods This consensus document was created by a multidisciplinary panel of 45 physicians with experience in the treatment of CAP in immunocompromised patients. The Delphi survey methodology was used to reach consensus. Results The panel focused on 21 questions addressing initial management strategies. The panel achieved consensus in defining the population, site of care, likely pathogens, microbiologic workup, general principles of empirical therapy, and empirical therapy for specific pathogens. Interpretation This document offers general suggestions for the initial treatment of the immunocompromised patient who arrives at the hospital with pneumonia

Appropriate age range for introduction of complementary feeding into an infant’s diet

Peer reviewedPublisher PD

Vaccine breakthrough hypoxemic COVID-19 pneumonia in patients with auto-Abs neutralizing type I IFNs

Life-threatening `breakthrough' cases of critical COVID-19 are attributed to poor or waning antibody response to the SARS- CoV-2 vaccine in individuals already at risk. Pre-existing autoantibodies (auto-Abs) neutralizing type I IFNs underlie at least 15% of critical COVID-19 pneumonia cases in unvaccinated individuals; however, their contribution to hypoxemic breakthrough cases in vaccinated people remains unknown. Here, we studied a cohort of 48 individuals ( age 20-86 years) who received 2 doses of an mRNA vaccine and developed a breakthrough infection with hypoxemic COVID-19 pneumonia 2 weeks to 4 months later. Antibody levels to the vaccine, neutralization of the virus, and auto- Abs to type I IFNs were measured in the plasma. Forty-two individuals had no known deficiency of B cell immunity and a normal antibody response to the vaccine. Among them, ten (24%) had auto-Abs neutralizing type I IFNs (aged 43-86 years). Eight of these ten patients had auto-Abs neutralizing both IFN-a2 and IFN-., while two neutralized IFN-omega only. No patient neutralized IFN-ss. Seven neutralized 10 ng/mL of type I IFNs, and three 100 pg/mL only. Seven patients neutralized SARS-CoV-2 D614G and the Delta variant (B.1.617.2) efficiently, while one patient neutralized Delta slightly less efficiently. Two of the three patients neutralizing only 100 pg/mL of type I IFNs neutralized both D61G and Delta less efficiently. Despite two mRNA vaccine inoculations and the presence of circulating antibodies capable of neutralizing SARS-CoV-2, auto-Abs neutralizing type I IFNs may underlie a significant proportion of hypoxemic COVID-19 pneumonia cases, highlighting the importance of this particularly vulnerable population